2-酮丁酸通过成纤维细胞生长因子受体3调节心肌细胞活力  

2-ketobutyric Acid Regulates Cardiomyocyte Viability Via Fibroblast Growth Factor Receptor 3

作　　者：方克宝 付美娇 徐方晶 张瑜[4] 孙美琪 马占兵[1] 何军[5] FANG Kebao;FU Meijiao;XU Fangjing;ZHANG Yu;SUN Meiqi;MA Zhanbing;HE Jun(Ningxia Medical University,Yinchuan 750004,China;Department of Geriatrics,Yantai Yuhuangding Hospital,Yantai 264001,China;Department of Critical Care Medicine,Yinchuan Hospital of Traditional Chinese Medicine Affiliated to Ningxia Medical University,Yinchuan 750004,China;Peking University First Hospital,Ningxia Women and Children,s Hospital,Yinchuan 750004,China;General Hospital of Ningxia Medical University,First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学,银川750004 [2]烟台毓璜顶医院老年医学科,烟台264001 [3]宁夏医科大学附属银川市中医医院重症医学科,银川750004 [4]北京大学第一医院宁夏妇女儿童医院,银川750004 [5]宁夏医科大学总医院,宁夏医科大学第一临床医学院,银川750004

出　　处：《宁夏医科大学学报》2025年第3期223-232,共10页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(2023AAC02070);宁夏重点研发计划项目(2021BEG03115)。

摘　　要：目的观察2-酮丁酸(2-KB)对心肌细胞生长活性的影响,并探讨其分子机制。方法分别将大鼠心肌细胞H9C2及人心肌细胞AC16各自设对照组和实验组,以2-KB与实验组细胞共孵育16 h。通过CCK-8实验、结晶紫染色法和细胞划痕实验检测心肌细胞活力。利用RNA测序筛选两种细胞中实验组与对照组共同的差异表达基因(differentially expressed genes,DEGs),通过CytoHubba筛选枢纽基因并经RT-qPCR验证,将验证后CytoHubba评分最高的基因进行基因干扰并检测干扰后心肌细胞活力的变化。结果CCK-8、结晶紫染色检测和划痕实验结果显示,实验组H9C2和AC16细胞活力均下降(P均<0.05)。RNA测序获得228个两种心肌细胞共有的DEGs,经CytoHubba筛选和RT-qPCR验证,获得4个关键枢纽基因,其中成纤维细胞生长因子受体3(fibroblast growth factor receptor 3,FGFR3)评分最高。FGFR3基因干扰实验结果显示,RNAi-FGFR3组心肌细胞活力较对照组和RNAi-vector组均降低(P均<0.05)。结论2-KB诱导心肌细胞活力降低、FGFR3表达上调,FGFR3表达上调可能是一种保护性机制。Objective To observe the effect of 2-ketobutyric acid(2-KB)on growth activity of myocardial cells and explore its molecular mechanisms.Methods The rat cardiomyocyte line H9C2 and the human cardiomyocyte line AC16 were divided into control group and experimental group respectively,with the experimental groups incubated with 2-KB for 16 h.The proliferation of cardiomyocytes was assessed using CCK-8 assay,crystal violet staining,and cell scratch assay.RNA sequencing was utilized to screen for common differentially expressed genes(DEGs)between the experimental and control groups in both types of cells.Key hub genes were identified using CytoHubba and validated by RT-qPCR.The highest-scoring gene from CytoHubba that was verified underwent gene interference,and changes in cardiomyocyte viability after interference were measured.Results Results from the CCK-8 assay,crystal violet staining,and scratch assays indicated that the viability of both H9C2 and AC16 cells in the experimental groups decreased(P all<0.05).A total of 228 common DEGs were obtained from RNA sequencing.After screening with CytoHubba and validating with RT-qPCR,four key hub genes were identified,among which fibroblast growth factor receptor 3(FGFR3)had the highest score.The results of FGFR3 gene interference experiments showed that the viability of cardiomyocytes in the RNAi-FGFR3 group was lower compared to the control group and the RNAi-vector group(P all<0.05).Conclusion 2-KB induces a decrease in cardiomyocytes viability and increase in FGFR3 expression.The increased FGFR3 expression may be a protective mechanism.

关 键 词：2-酮丁酸 心肌细胞活力 RNA测序 成纤维细胞生长因子受体3 

分 类 号：R541.6[医药卫生—心血管疾病]